Scientia Agricultura Sinica ›› 2021, Vol. 54 ›› Issue (14): 2931-2940.doi: 10.3864/j.issn.0578-1752.2021.14.001

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Production of High Linoleic Acid Rice by Genome Editing

CHEN XiaoJun1(),WU KaiShen2,HUI Jian1,BAI HaiBo1,MA SiShuang1,LI JingYu2   

  1. 1Agricultural Biotechnology Center, Ningxia Academy of Agriculture and Forestry Sciences/Key Lab of Agricultural Biotechnology of Ningxia, Yinchuan 750002
    2College of Biological Science & Engineering, North Minzu University, Yinchuan 750021
  • Received:2020-11-02 Accepted:2021-01-29 Online:2021-07-16 Published:2021-07-26

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Abstract:

【Objective】 Linoleic acid is one of essential fatty acid for human, which can accelerate the metabolism and decomposition of fat in the body, reduce the accumulation of cholesterol on the wall of blood vessels, effectively prevent the occurrence of arteriosclerosis, improve human immunity, promote bone development, improve memory and prevent brain function degradation. Rice bran is a by-product of rice processing, and is an edible oil with high nutritional value. In this paper, we precisely edited the gene of ω-3 fatty acid desaturase, which is the key enzyme controlling fatty acid synthesis pathway. It can enrich the upstream linoleic acid, while closing the synthesis pathway of downstream products. 【Method】There are two copies of the key enzyme ω-3 fatty acid desaturase gene ( OsFAD3) in the fatty acid synthesis pathway in rice, which are located on chromosome 11 and chromosome 12, and their cDNA homology is 97.32%. According to the basic principles of gene editing, the specificity of edit depends on the characteristics of the guide RNA (gRNA). In this study, two gRNAs were designed and synthesized in the exon region of two homologous genes, and were constructed into editing vectors respectively. The local receptor material (Fuyuan 4) was successfully transformed by Agrobacterium mediated method. The editing efficiency and genotype of the two sites were analyzed. At the same time, the main agronomic characters of OsFAD3 double mutant were measured, such as grain width and grain length. The contents of 37 fatty acids were figured by GC-MS in the grains of OsFAD3double mutants. 【Result】The results showed two homozygous editing materials were obtained, and other editing materials with different genotypes were obtained. Compared with the control materials, the main agronomic traits such as grain width and grain length of OsFAD3 double mutant materials had no significant changes, however, the relative content of linoleic acid in rice increased by 3.36%. 【Conclusion】One gRNA vector knock out two ω-3 fatty acid desaturase genes at the same time, which improves the relative content of linoleic acid in grains without changing the main agronomic traits of seeds.

Key words: rice, gene edit, high linoleic, rice bran

Fig. 1

The pathway of fatty acid synthesis in rice[4] "

Fig. 2

Structure comparison of OsFAD3.1 and OsFAD3.2 "

Fig. 3

Design of gRNA1 target site"

Fig. 4

Plant gene editing vector"

Fig. 5

Sequence gene editing vector"

Fig. 6

Simultaneous editing of OsFAD3.1 and OsFAD3.2 A, B: FAD3-S1-PC1300 editing plant; C, D: FAD3-S2-PC1300 editing plant. A, C: The target of OsFAD3.1; B, D: The target of OsFAD3.2.Arrows indicate editing position "

Fig. 7

The sequence of editing plants A: The sequence of editing plants with FAD3-S1-PC1300; B: The sequence of editing plants with FAD3-S2-PC1300"

Fig. 9

Distribution of nine genotypes in the progeny of gene editing"

Fig. 10

Agronomic trait of grain in OsFAD3 double mutant A: Thousand grain weight; B: Average grain length; C: Average grain width"

Fig. 11

Comparison of fatty acid content"

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